Molecular Insights into the Thermal Stability of mAbs with Variable-Temperature Ion-Mobility Mass Spectrometry
Autor: | David P. Humphreys, Richard J. K. Taylor, Rachel A. Garlish, Shirley Jane Peters, Perdita E. Barran, Kamila J. Pacholarz, Alistair James Henry |
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Rok vydání: | 2015 |
Předmět: |
0301 basic medicine
Protein Conformation Ion-mobility spectrometry Mass spectrometry Biochemistry Mass Spectrometry 03 medical and health sciences Protein structure Molecule Thermal stability Immunoglobulin Fragments Molecular Biology Chromatography Protein Stability Chemistry Organic Chemistry Temperature Antibodies Monoclonal Protein engineering 030104 developmental biology Immunoglobulin G Biophysics ion mobility-mass spectrometry Molecular Medicine Chemical stability |
Zdroj: | Pacholarz, K, Peters, S, Garlish, R, Henry, A, Taylor, R, Humphreys, D & Barran, P 2016, ' Molecular Insights into the Thermal Stability of mAbs with Variable-Temperature Ion-Mobility Mass Spectrometry ', ChemBioChem: a European journal of chemical biology . https://doi.org/10.1002/cbic.201500574 |
ISSN: | 1439-4227 |
Popis: | The aggregation of protein-based therapeutics such as monoclonal antibodies (mAbs) can affect the efficacy of the treatment and can even induce effects that are adverse to the patient. Protein engineering is used to shift the mAb away from an aggregation-prone state by increasing the thermodynamic stability of the native fold, which might in turn alter conformational flexibility. We have probed the thermal stability of three types of intact IgG molecules and two Fc-hinge fragments by using variable-temperature ion-mobility mass spectrometry (VT-IM-MS). We observed changes in the conformations of isolated proteins as a function of temperature (300-550 K). The observed differences in thermal stability between IgG subclasses can be rationalized in terms of changes to higher-order structural organization mitigated by the hinge region. VT-IM-MS provides insights into mAbs structural thermodynamics and is presented as a promising tool for thermal-stability studies for proteins of therapeutic interest. |
Databáze: | OpenAIRE |
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